Methods of controlling pests using terpendoles

ABSTRACT

The present invention is directed to a method of controlling a pest comprising applying an effective amount of one or more terpendoles to the pest or an area in need of pest control. The present invention is further directed to a method of controlling a pest comprising applying an effective amount of one or more compounds having the following chemical structure 
                         
to the pest or an area in need of pest control.

FIELD OF THE INVENTION

The present invention relates to a method of controlling a pestcomprising applying an effective amount of one or more terpendoles tothe pest or an area in need of pest control.

The present invention further relates to a method of controlling a pestcomprising applying an effective amount of one or more compounds havingthe following chemical structure

to the pest or an area in need of pest control.

BACKGROUND OF THE INVENTION

Arthropod pests are one of the major threats to human welfare and exertcontinued stress on the food supply via herbivory, fouling and diseasetransmission. Synthetic insecticides played a significant role and, inmany ways, ushered in modern agriculture and pest control. However,there is increasing pressure from the public and from regulatoryagencies to reduce or eliminate the exclusive use of synthetic chemicalin the control of agricultural arthropod pests. The widespread use ofavailable insecticides has resulted in the development of resistantinsect populations. Insecticide resistance is a complex phenomenonmanifested in a diverse array of physiological and/or behavioralmechanisms. Major mechanisms that are responsible for the development ofinsecticide resistance are metabolic detoxification, target sitemutation, reduced cuticular penetration and behavioral avoidance. Novelclasses of natural insecticides are needed to combat the ever-increasingnumber of resistant insect species and populations.

Indole-diterpenes are a natural, structurally diverse group of secondarymetabolites with a common cyclic diterpene backbone derived fromgeranylgeranyl diphosphate and an indole group derived fromindole-3-glycerol phosphate. Terpendoles are a specific class ofindole-diterpene alkaloids produced by fungi that were first discoveredby Huang et al. when screening for acyl-CoA:cholesterol acyltransferaseinhibitors. Huang et al. Terpendoles, Novel ACAT inhibitors produced byAlbophoma yamanashiensis I. production, isolation and biologicalproperties, J Antibiot (Tokyo). 1995 January, 48(1), 1-4. Specifically,Huang et al. discovered terpendoles A-D. Since that time, 10 additionalterpendoles have been described in the literature and termed terpendolesE-M. To date, terpendoles have been further discovered to inhibit motoractivation of mitotic kinesin Eg5. See, Nakazawa J. et al., A novelaction of terpendole E on the motor activity of mitotic Kinesin Eg5,Chem Biol, 2003 February, 10(2), 131-137. Due to its role in inhibitionmitotic kinesin, terpendole E is currently being researched as apossible anti-cancer drug. However, there is no known application ofterpendoles that would benefit humans.

Given that there is currently no known use for terpendoles and alsogiven that resistance is problematic for insect pest populations, weassert that there is a need in the art for novel uses of terpendoles asnatural pesticides.

SUMMARY OF THE INVENTION

The present invention is directed to a method of controlling a pestcomprising applying an effective amount of one or more terpendoles tothe pest or an area in need of pest control.

The present invention is further directed to a method of controlling apest comprising applying an effective amount of one or more compoundshaving the following chemical structure

to the pest or an area in need of pest control.

DETAILED DESCRIPTION OF THE INVENTION

According to the present invention terpendoles are an effectivepesticide. This discovery is unexpected as no known use for terpendoleshad previously been discovered.

As used herein “terpendole A” refers to the following structure

with CAS number 156967-64-5.

As used herein “terpendole B” refers to the following structure

with CAS number 156967-67-8.

As used herein “terpendole C” refers to the following structure

with CAS number 156967-65-6.

As used herein “terpendole D” refers to the following structure

with CAS number 156967-66-7.

As used herein “terpendole E” refers to the following structure

with CAS number 167427-23-8.

As used herein “terpendole F” refers to the following structure

with CAS number 167427-24-9.

As used herein “terpendole G” refers to the following structure

with CAS number 167427-25-0.

As used herein “terpendole H” refers to the following structure

with CAS number 156967-69-0.

As used herein “terpendole I” refers to the following structure

with CAS number 167612-17-1.

As used herein “terpendole J” refers to the following structure

with CAS number 167427-26-1.

As used herein “terpendole K” refers to the following structure

with CAS number 167427-27-2.

As used herein “terpendole L” refers to the following structure

with CAS number 167612-18-2.

As used herein “terpendole M” refers to the following structure

with CAS number 222400-32-0.

As used herein “terpendole N” refers to the following structure

As used herein “terpendole O” refers to the following structure

As used herein “terpendole P” refers to the following structure

The one or more terpendoles of the present invention can be applied byany convenient means. Those skilled in the art are familiar with themodes of application including but not limited to, spraying, brushing,soaking, in-furrow treatments, drip irrigation, granule application,seed treatment, pressurized liquids (aerosols), fogging orside-dressing.

As used herein, “to control” a pest or “controlling” pest(s) refers tokilling, incapacitating, repelling, or otherwise decreasing the negativeimpact of the pest on plants or animals to a level that is desirable tothe grower, applicator or user.

As used herein, “composition” refers to one or more active ingredientsin a carrier. The carrier may be a liquid, a semi-solid, a solid or agas and may contain additional ingredients. For example, a fermentationbroth is a suitable carrier for the present invention.

As used herein, “an area in need of pest control” refers to any areathat the pest is present during any life stage. One environment likelyto be treated by the methods of the present invention includes theplants that the pest is living on and/or the surrounding soil. Thepest's environment may also include an area where plants are grown,harvested, or in gardens, fields, greenhouses, or other buildings, andvarious indoor surfaces and structures, such as furniture includingbeds, and furnishings including books, clothing, etc.

As used herein, all numerical values relating to amounts, weightpercentages and the like are defined as “about” or “approximately” eachparticular value, namely, plus or minus 10%. For example, the phrase“about 5,000 parts per million” is to be understood as “from 4,500 to5,500 parts per million.” Therefore, amounts within 10% of the claimedvalues are encompassed by the scope of the claims.

The term “effective amount” means the amount of the one or moreterpendoles that will control the target pest. The “effective amount”will vary depending on the terpendole concentration, the type of pest(s)being treated, the severity of the pest infestation, the result desired,and the life stage of the pest during treatment, among other factors.Thus, it is not always possible to specify in advance an exact“effective amount.” However, an appropriate “effective amount” in anyindividual case may be determined by one of ordinary skill in the art.

The articles “a,” “an” and “the” are intended to include the plural aswell as the singular, unless the context clearly indicates otherwise.For example, the methods of the present invention are directed tocontrolling “pest” but this can include control of a multiple pests(such as a more than one insect or more than one insect species or morethan one mite or more than one mite species).

In one embodiment, the present invention is directed to a method ofcontrolling a pest comprising applying an effective amount of one ormore terpendoles to the pest or an area in need of pest control.

In a preferred embodiment, the pest is an arthropod pest, morepreferably an insect or a mite. As used herein, “arthropod” refers topests that belong to the Phylum Arthropoda. As used herein, “insect”refers to pests that belong to the Class Insecta. As used herein, “mite”refers to pests that belong to the Subclass Acari of the ClassArachnida.

In another preferred embodiment, the pest is an aphid. As used herein,“aphid” refers to pests that belong to the Family Aphididae. Exemplaryaphids include cotton aphid (Aphis gossypii), foxglove aphid(Aulacorthum solani), cabbage aphid (Brevicoryne brassicae),birdcherry-oat aphid (Rhopalosiphum padi) and green peach aphid (Myzuspersicae).

In another preferred embodiment, the pest is a lepidopteran. As usedherein, “lepidopteran” refers to pests that belong to the OrderLepidoptera including moths and their larval stages. Exemplarylepidopterans include diamondback moth (Plutella xylostella) and commoncutworm (Spodoptera litura).

In another preferred embodiment, the pest is a thrips. As used herein,“thrips” refers to pests that belong to the Family Thripidae. Exemplarythrips include western flower thrips (Frankliniella occidentalis).

In another preferred embodiment, the pest is a whitefly. As used herein,“whitefly” refers to pests that belong to the Family Aleyrodidae.Exemplary whiteflies include silverleaf whitefly and tobacco whitefly(Bemisia tabaci).

In another preferred embodiment, the pest is a planthopper. As usedherein, “planthopper”, refers to pests that belong to the InfraorderFulgoromorpha of the Order Hemiptera. Exemplary planthoppers includebrown rice planthopper (Nilaparvata lugens).

In another preferred embodiment, the pest belongs to the infraclassNeoptera of the Class Insecta, the Subfamily Aphidinae of the FamilyAphididae or the Parvorder Heteroneura of the Family Lepidoptera.

In another preferred embodiment, the area in need of pest control is aplant.

In another preferred embodiment, the area in need of pest control is anarea that includes but is not limited to, where crops are grown,harvested, stored, processed, packed or shipped.

Terpendoles include, but are not limited to, compounds having thefollowing chemical structure

wherein:R¹ and R² are each independently H or 2-methyl-2-butene;R³ and R⁴ are each independently H or —OH or R³ and R⁴ are takentogether to form an epoxide bridge;R⁵ is CH₃, —CH₂—OH, or —CH═O and R⁶ is —OH or R⁵ and R⁶ are takentogether to form an epoxide bridge;R⁷ and R¹⁰ are each H or absent, wherein if R⁷ and R¹⁰ are absent thencarbon atoms adjacent to R⁷ and R¹⁰ are double bonded; andR⁸ is H or —OH and R⁹ is —C—(CH₃)₂—O—CH₂—CH═C—(CH₃)₂, —C—(CH₃)₂—OH or R⁸and R⁹ taken together form

In a preferred embodiment, the one or more terpendoles are selected fromthe group consisting of terpendole A, terpendole B, terpendole C,terpendole D, terpendole E, terpendole F, terpendole G, terpendole H,terpendole I, terpendole J, terpendole K, terpendole L, terpendole M,terpendole N, terpendole O and terpendole P, more preferably the one ormore terpendoles is selected from terpendole A, terpendole C, terpendoleK, terpendole N, terpendole O and terpendole P.

The one or more terpendoles of the present invention may be applied at aconcentration from about 1 to about 10,000 parts per million (“ppm”),preferably from about 4 to about 5,000 parts per million, morepreferably from about 4 to about 100 ppm.

In another embodiment, the one or more terpendoles are an activeingredient in a composition.

Other components that enhance the biological activity or application ofthese ingredients may optionally be included.

The following example is intended to illustrate the present inventionand to teach one of ordinary skill in the art how to use the one or moreterpendoles of the invention and is not intended to be limiting in anyway.

EXAMPLES Example 1 Isolation of Terpendoles from Tolypocladium inflatumFermentation Broth

Fermentation broth of Tolypocladium inflatum was extracted with ethylacetate. Evaporation of the ethyl acetate solution gave a dark thicksyrup. This material was re-dissolved in an appropriate amount of ethylacetate-methanol (2:1) mixture, filtered and combined with anappropriate amount of silica gel. The mixture was then evaporated on arotary evaporator until dry. The resulting solid was chromatographed ona silica gel column with a slow gradient of ethyl acetate in hexane. Thefractions containing terpendoles, as determined with liquidchromatography mass spectrometry (LC-MS) analysis, were combined andevaporated. The resulting material was re-purified with silica gelchromatography or high-pass liquid chromatography, as appropriate toisolate the individual terpendoles.

Of the terpendoles found, terpendole A, terpendole C, terpendole J andterpendole K were previously described in literature. These compoundswere identified by comparing molecular weight, ¹H nuclear magneticresonance (“NMR”) and ¹³C NMR spectra with the reported spectra (Huang,et. al, J. Antibiotics, 1995, 48, 5; Tomoda, et al., J. Antibiotics,1995, 48, 793), as well as two-dimensional NMR COSY, NOESY, HSQC andHMBC. Spectral data of terpendole N, terpendole O and terpendole P havenot previously been reported. The structural determination of thesethree compounds is described in Examples 2-4, below. The relative stereoconfigurations of these compounds were established based on thestructures of known terpendoles.

Example 2 Structure Determination of Terpendole N

Mass spectrum of this compound indicates a molecular weight of 533. ¹HNMR (400 MHz, CDCl₃): δ7.73 (s, 1H, NH), 7.48 (d, 1H), 7.29 (d, 1H),7.07 (m, 2H), 5.39 (dd, 1H), 4.58 (d, 1H), 4.10 (d, 1H), 3.92 (s, 1H),3.90 (d, 1H), 3.20 (d, 1H), 2.88 (d, 1H), 2.73-2.71 (m, 2H),2.47-2.43(m, 1H), 2.02-1.80 (m, 2H), 1.69-1.46 (m, 1H), 1.37-1.99 (m,17H, including CH₃ at 1.35, 1.34, 1.33, 1.31 and 1.24), 1.13 (s, 3H).¹³C NMR (100 MHz, CDCl₃): δ151.57, 144.40, 139.86, 125.18, 120.60,119.74, 118.59, 117.65, 111.47, 106.19, 95.61, 76.24, 75.22, 73.13,71.09, 64.87, 62.58, 60.14, 57.86, 50.70, 49.94, 43.78, 30.58, 27.72,27.26, 24.52, 20.55, 20.00, 19.16, 16.50, 16.21. In comparison with thespectrum of terpendole A, this compound has one extra peak in thearomatic/vinyl region of the ¹H NMR at 5.39 ppm (dd) and two extra sp2carbon signals at 144.40 (C/CH₂ type) and 106.19 (CH/CH₃ type). Thesedata confirm that this compound has a tri-substituted carbon-carbondouble bond. Direct evidence for assigning this double bond at C6═C7 wasfound in the 2D HMBC spectrum, with the C6 vinyl proton at 5.39 ppmshowing strong interaction with neighboring C4, C12 and C7. Other NMRspectra, including 2D COSY, NOESY and HSQC were used to assign theproton and carbon signals.

Example 3 Structure Determination of Terpendole O

Mass spectrum of this compound indicates a molecular weight of 603. ¹HNMR (400 MHz, CDCl₃): δ7.73 (s, 1H, NH), 7.13 (d, 1H), 7.00 (t, 1H),6.84 (d, 1H), 5.40 (m, 1H), 4.62 (d, 1H), 4.34 (t, 1H), 3.86 (d, 1H),3.67 (s, 1H), 3.61-3.59 (m, 3H), 2.88 (d, 1H), 2.83-2.50 (m, 3H), 2.26(m, 1H), 1.94-1.90 (m, 1H), 1.756 (s, 3H), 1.754 (s, 3H), 1.67-1.53 (m,3H), 1.38-1.23 (m, 15H, including CH₃ at 1.33, 1.32, 1.30 and 1.27),1.21 (s, 3H), 1.13 (s, 3H). ¹³C NMR (100 MHz, CDCl₃): δ151.06, 139.69,133.11, 131.81, 124.51, 123.71, 120.96, 119.00, 117.15, 109.28, 95.49,78.19, 75.06, 71.62, 71.17, 71.10, 67.90, 62.59, 60.97, 57.78, 50.39,50.16, 42.45, 32.02, 30.38, 29.03, 28.09, 28.04, 27.46, 25.79, 24.57,20.55, 19.17, 18.86, 17.97, 16.46, 15.97.

Evidence for the presence of a (CH₃)₂C═CHCH₂— moiety was provided bystrong coupling of the vinyl proton at 5.40 ppm with the benzylic CH₂ at3.60 ppm and with the allylic methyl groups at 1.756 and 1.754 ppm inthe 2D-COSY spectrum. Further evidence was provided by the stronginteraction of the benzylic CH₂ at 3.60 ppm with C38 at 123.71 ppm andC39 at 131.81 ppm in 2D-HMBC spectrum. That the (CH₃)₂C═CHCH₂— moiety isattached to C20 was derived from the interaction of the benzylic CH₂ at3.60 ppm with C21 at 119.00 ppm. Other NMR spectra, including 2D COSY,NOESY and HSQC were used to assign the proton and carbon signals.

Example 4 Structure Determination of Terpendole P

Mass spectrum of this compound indicates a molecular weight of 603. ¹HNMR (400 MHz, CDCl₃): δ7.63 (s, 1H, NH), 7.33 (d, 1H), 7.01 (s, 1H),6.90 (d, 1H), 5.36 (m, 1H), 4.62 (d, 1H), 4.33 (t, 1H), 3.85 (d, 1H),3.67 (s, 1H), 3.59 (d, 1H), 3.41 (d, 2H), 2.88 (d, 1H), 2.81-2.60 (m,4H), 2.40-2.22 (m, 2H), 1.739 (s, 3H), 1.736 (s, 3H), 1.67-1.41 (m, 3H),1.38-1.23 (m, 11H, including CH₃ at 1.32, 1.31, and 1.30), 1.25 (s, 3H),1.21 (s, 3H), 1.10 (s, 3H). ¹³C NMR (100 MHz, CDCl₃): δ151.16, 140.24,134.42, 131.74, 124.23, 123.33, 120.63, 118.54, 117.52, 110.78, 95.48,78.16, 75.05, 71.62, 71.16, 71.09, 67.89, 62.75, 60.96, 57.78, 50.67,49.98, 42.44, 34.54, 30.40, 28.08, 28.03, 27.44, 27.23, 25.77, 24.57,20.59, 19.17, 18.82, 17.81, 16.46, 15.94.

Evidence for the presence of a (CH₃)₂C═CHCH₂— moiety was provided bystrong coupling of the vinyl proton at 5.36 ppm with the benzylic CH₂ at3.41 ppm and with allylic methyl groups at 1.739 and 1.736 ppm in the2D-COSY spectrum. Further evidence was provided by the stronginteraction of the benzylic CH₂ at 3.41 ppm with C38 at 124.23 ppm andC39 at 131.74 ppm in 2D-HMBC spectrum. Unlike in terpendole O, thebenzylic CH₂ at 3.41 ppm also has strong interaction with C22 at 134.42ppm and C23 at 110.79 ppm in the 2D HMBC spectrum, lending support toattachment of (CH₃)₂C═CHCH₂— moiety to C22. Other NMR spectra, including2D COSY, NOESY and HSQC were used to assign the proton and carbonsignals.

Example 5 Insecticidal Activity of Terpendoles

Method

Biological efficacy assays utilizing terpendoles were conducted on greenpeach aphids (Myzus persicae). Specifically, terpendole A, terpendole C,terpendole J, terpendole K, terpendole N, terpendole O and terpendole Pwere each separately applied in serial dilutions to determine theconcentration at which 50% control of green peach aphids occurred over aperiod of 24-48 hours (“LC₅₀”). Radish leaves infested with green peachaphids were treated, top and bottom, with a total of 500 μl of anaerosolized terpendole dilution. Treated, aphid infested leaves wereheld in greenhouse conditions and evaluated for efficacy at 24 and 48 h.

Results

As seen in Table 1 below, each of terpendole A, terpendole C, terpendoleJ, terpendole K, terpendole N, terpendole O and terpendole P,effectively controlled green peach aphids. Specifically, terpendoleshave an LC₅₀ as low as 4 parts per million (“ppm”) against Myzuspersicae.

TABLE 1 LC₅₀ of Terpendoles on Green Peach Aphids Compound LC₅₀ (ppm)Terpendole A 7 Terpendole C 20 Terpendole J 5,183 Terpendole K 44Terpendole N 4 Terpendole O 105 Terpendole P 67

Example 6 Insecticidal Activity of Terpendole C

Method

A bioefficacy assay utilizing terpendole C was conducted on thelepidopterans: diamondback moth (Plutella xylostella) and common cutworm(Spodoptera litura): the whitefly, tobacco whitefly (Bemisia tabaci) thethrips, western flower thrips (Frankliniella occidentalis); theplanthopper, brown rice planthopper (Nilaparvata lugens); the aphids,cotton aphid (Aphis gossypii), foxglove aphid (Aulacorthum solani),cabbage aphid (Brevicoryne brassicae), birdcherry-oat aphid(Rhopalosiphum padi), green peach aphid (Myzus persicae); and the mite,two-spotted spider mite (Tetranychus urticae). Specifically, terpendoleC was diluted with acetone and 5% Tween® 20 and finally diluted withwater to prepare spray solutions. Terpendole C was then applied toplants as a foliar spray at 500, 200, 50, 12.5, 3.1, 1, 0.8 and/or 0.2ppm and/or as a soil treatment at 2,500, 1,000, 250, 63, 16, 4 and/or 1ppm. Pests were then placed on the plant or in the soil. Certain pestswere at particular life cycle stages during application as indicated inTables 2 and 3, below. Further, mortality and appetite suppression wererecorded at 6, 7 and/or 13 days after treatment (“DAT”) and are reportedin Tables 2 and 3, below, respectively. A score of 0 does not indicateno mortality or no feeding suppression but instead a range of mortalityor feeding suppression less than 29% or 5%, respectively.

TABLE 2 Mortality Rates of Various Insects Target Pest Plant Stage DAT500 200 50 12.5 3.1 1 0.8 0.2 Diamondback Cabbage 3rd instar 6 4 3 1 1 —— — — moth Common Cabbage 4th instar 6 0 0 0 0 — — — — cutworm TobaccoCabbage 1st instar 7 0 0 0 0-2 1 1 0 — whitefly Western Kidney 1st-2ndinstar 6 0 0 0 0 0 — 0 0 flower Bean thrips Brown rice Rice 3rd-4thinstar 6 0 0 0 0-1 0 — 0 0 plant hopper Cotton aphid Cucumber All 6 4 43 3-4 3 — 2 0 Foxglove Kidney Adult 6 2 1 0 0 — — — — aphid Bean Cabbageaphid Cabbage All 6 — 4 4 4 2 — 2 — Bird Wheat All 6 — 3 2 1 0 — 0 —cherry-oat aphid Green peach Cabbage All 6 — 4 3 2 1 — 0 — aphidTwo-spotted Kidney All 7 0 0 0 0 — — — — spider mite Bean 13  0 0 0 0 —— — — 0 = about 29% mortality or less 1 = about 50% mortality 2 = about75% mortality 3 = about 90% mortality 4 = 100% mortality

TABLE 3 Appetite Suppression of Various Insects Pest Plant Target StageDAT 500 200 50 12.5 Diamondback Cabbage 3rd instar 6 4 3 0 0 moth CommonCabbage 4th instar 6 4 4 4 4 cutworm Western Kidney 1st-2nd 6 3-4 3-43-4 3-4 flower thrips Bean instar Foxglove Kidney Adult 6 2 0 0 0 aphidBean Two-spotted Kidney All 7 0 0 0 0 spider mite Bean 13  0 0 0 0 0 =no to little suppression 1 = about 10% suppression 2 = about 25%suppression 3 = about 60% suppression 4 = 70-100% suppressionResults

As seen in Table 2 above, terpendole C effectively controlled the aphidsincluding: cotton aphids, foxglove aphids, cabbage aphids,birdcherry-oat aphids and green peach aphids. Further, terpendole Ceffectively controlled the lepidopterans, including diamondback moth andcommon cutworm, either by killing the larvae or suppressing the appetiteof the larvae. Finally, terpendole C significantly controlled thewhitefly and the thrips.

What is claimed is:
 1. A method of controlling an aphid comprisingapplying an effective amount of one or more terpendoles to the pest oran area in need of aphid control, wherein each of the one or moreterpendoles have the following chemical structure

wherein: R¹ and R² are each independently H or 2-methyl-2-butene; R³ andR⁴ are each independently H or —OH or R³ and R⁴ are taken together toform an epoxide bridge; R⁵ is CH_(3,) —CH₂—OH, or —CH═O and R⁶ is —OH orR⁵ and R⁶ are taken together to form an epoxide bridge; R⁷ and R¹⁰ areeach H or absent, wherein if R⁷ and R¹⁰ are absent then carbon atomsadjacent to R⁷ and R¹⁰ are double bonded; and R⁸ is H or —OH and R⁹ is—C—(CH₃)₂—O—CH₂—CH═C—(CH₃)₂, —C—(CH₃)₂—OH or R⁸ and R⁹ taken togetherform


2. The method of claim 1, wherein the aphid is selected from the groupconsisting of cotton aphid (Aphis gossypii), foxglove aphid (Aulacorthumsolani), cabbage aphid (Brevicoryne brassicae), birdcherry-oat aphid(Rhopalosiphum padi) and green peach aphid (Myzus persicae).
 3. Themethod of claim 1, wherein the effective concentration is from about 1to about 10,000 parts per million.
 4. The method of claim 1, wherein theeffective concentration is from about 4 to about 5,000 parts permillion.
 5. The method of claim 1, wherein the one or more terpendolesare selected from the group consisting of terpendole A, terpendole B,terpendole C, terpendole D, terpendole E, terpendole F, terpendole G,terpendole H, terpendole I, terpendole J, terpendole K, terpendole L,terpendole M, terpendole N, terpendole O and terpendole P.
 6. The methodof claim 1, wherein the one or more terpendoles is selected fromterpendole A, terpendole C, terpendole K, terpendole N, terpendole O andterpendole P.